Funcionalização do polietilenoglicol metil éter via reação de bromoacetilação: ativação, caracterização por MALDI-TOF e mecanismo de reação / Poly(ethylene glycol) methyl ether functionalization by bromoacetylation reaction: Activation, characterization by MALDI-TOF and reaction mechanism / Funcionalización de polietilenglicol metil éter mediante reacción de bromoacetilación: activación, caracterización por MALDI-TOF y mecanismo de reacción

Resumo A PEGuilação, reação química de conjugação com a molécula de polietilenoglicol (PEG) ou polietilenoglicol metil éter (mPEG), tem sido amplamente aplicada pelas indústrias farmacêuticas como estratégia de melhoria das propriedades farmaco-cinéticas de compostos bioativos. O PEG é um polímero que possui um esqueleto de poliéter quimicamente inerte e que apresenta grupos hidroxilas (-OH) em suas extremidades. Assim, o PEG para tornar-se apto como reagente de conjugação deve ser ativado com um grupo funcional que seja reativo. Nesse sentido, a bromoaceti-lação apresenta-se como uma alternativa para a funcionalização do PEG. Portanto, nesse trabalho objetivamos descrever em detalhes os procedimentos e o mecanismo de reação envolvida na funcionalização do mPEG, através da reação de bromoacetilação. Além do mais, estudamos a aplicação do MALDI-ToF para a caracterização do produto ativado. Após a bromoacetilação, por um procedimento adaptado, obteve-se o bromoacetil-mPEG-éster, com rendimento bruto de 56,78%. Análises posteriores, por espectrometria de massas por MALDI-ToF, possibilitaram identificar e caracterizar o produto bromoacetilado. Entre as condições de reação, o controle de temperatura (-10 °C a 0 °C) mostrou-se eficaz favorecendo a adição nucleofílica essencial à bromoacetilação. Assim, concluímos que o controle da baixa temperatura reacional é um fator chave para o favorecimento da adição nucleofílica à carbonila e, portanto, essencial na obtenção do mPEG funcionalizado via bromoacetilação. Estudos posteriores serão necessários, no entanto, para confirmar se o mPEG esterificado, nessas condições, poderá ser utilizado na conjugação com moléculas de natureza proteica ou peptídica, por meio de substituição nucleofílica bimolecular.

SUMMARY PEGylation, a chemical reaction of conjugation with the polyethylene glycol molecule (PEG), has been widely applied by the pharmaceutical industries as a strategy to improve the pharmacokinetic properties of bioactive compounds. PEG is a polymer that has a chemically inert polyether backbone and hydroxyl groups (-OH) at its ends. Thus, PEG to become fit as a reagent for conjugation must be activated with a functional group that is reactive. In this sense, bromoacetylation presents itself as an alternative for the functionalization of PEG. Therefore, in this study we aim to describe in detail the procedures and reaction mechanism involved in the functionalization of mPEG through the bromoacetylation reaction. In addition, we used the spectrometric technique, by MALDI-ToF, for the characterization of the activated product. After applying an adapted bromoacetylation procedure, bromoacetyl-mPEG-ester was obtained with a yield of 56.78%. Subsequent analyzes of MALDI-ToF mass spectrometry were able to correctly identify and characterize the bromoacety-lated product. Among the reaction conditions, temperature control (from -10 °C to 0 °C) was effective in favoring the essential nucleophilic addition to bromoacetylation. Thus, we conclude that the control of the low reaction temperature is a key factor in favoring the nucleophilic addition to carbonyl and, therefore, obtaining a favorable conversion to functionalized PEG via bromoacetylation. Further studies, however, will be necessary to confirm whether PEG esterified with these conditions can be used in conjunction with molecules of a protein or peptide nature by means of bimolecular nucleophilic substitution.

RESUMEN La PEGilación, una reacción química de conjugación con la molécula de polietilenglicol (PEG), ha sido ampliamente aplicada por las industrias farmacéuticas como una estrategia para mejorar las propiedades farmacocinéticas de los compuestos bioactivos. El PEG es un polímero formado por un esqueleto de poliéter químicamente inerte con grupos hidroxilo (-OH) en sus extremos. Por lo tanto, para usar el PEG como reactivo de conjugación debe activarse con un grupo funcional que sea reactivo. En este sentido, la bromoacetilación es una alternativa para la funcionalización de PEG. De esta manera, en este trabajo nuestro objetivo es describir en detalle los procedimientos y el mecanismo de reacción involucrados en la funcionalización de PEG a través de la reacción de bromoacetilación. Además, estudiamos la aplicación de MALDI-ToF para la caracterización del producto activado. Después de aplicar un procedimiento de bromoacetilación adaptado, se obtuvo bromoacetil-mPEG-éster con un rendimiento bruto de 56,78%. Los análisis posteriores de espectrometría de masas por MALDI-ToF pudieron identificar y caracterizar correctamente el producto bromoacetilado. Entre las condiciones de reacción, el control de la temperatura (desde -10 °C hasta 0 °C) fue eficaz para favorecer la adición nucleofílica esencial a la bromoacetilación. Así, concluimos que el control de la baja temperatura de reacción es un factor clave para favorecer la adición nucleofílica al carbonilo y, por lo tanto, esencial para obtener el mPEG funcionalizado mediante la bromoacetilación. Sin embargo, serán necesarios más estudios para confirmar si el mPEG esterificado en estas condiciones puede usarse junto con moléculas de naturaleza proteica o peptídica por medio de la sustitución nucleófila bimolecular.

Addition of chlorogenic acid and caffeine during the processing of cooled boar semen / Adição de ácido clorogênico e cafeína durante o processamento do sêmen suíno resfriado

A study was conducted to evaluate the effect of chlorogenic acid (ChA) added pre-cooling and its combination with caffeine added during warming on cooled-stored boar semen parameters. Ten ejaculates were diluted in commercial extender with or without 4.5mg/ml ChA and stored at 15°C. After 0, 24 and 72 hours of storage, aliquots of these doses were taken and incubated at 37°C in the presence or absence of 8.0mM caffeine. Semen quality was evaluated after 10 and 120 minutes of incubation. The ChA increased (P <0.01) the sperm motility, viability, acrosomal integrity and the percentage of spermatozoa with high mitochondrial activity (PMHA), however, decreased (P <0.01) the malondialdehyde (MDA) concentration. Caffeine increased (P<0.05) the sperm motility, viability, PMHA and the MDA concentration and reduced (P <0.05) the acrosome integrity. When associated (ChA+caffeine), there was an increase (P <0.05) in sperm motility and viability, PMHA and acrosome integrity. The addition of ChA to the dilution medium improves the quality of the swine inseminating doses. The addition of caffeine during re-warming is only recommended when the semen is stored for prolonged periods (72h), and the inseminating dose should be used immediately after its addition.(AU)

O objetivo deste estudo foi avaliar os efeitos da adição de ácido clorogênico (ChA) antes do resfriamento e sua combinação com cafeína adicionada durante o reaquecimento sobre a qualidade do sêmen suíno resfriado. Dez ejaculados foram diluídos em diluidor comercial com adição ou não de 4,5mg/mL de ChA e armazenados a 15°C. Após zero, 24 e 72 horas de armazenamento, 10mL foram retirados e incubados a 37°C na presença ou ausência de 8,0mM de cafeína. A qualidade seminal foi avaliada após 10 e 120 minutos de incubação. O ChA aumentou (P<0,01) a motilidade, a viabilidade, a integridade acrosomal e a porcentagem de espermatozoides com alta atividade mitocondrial (PMHA), entretanto diminuiu (P<0,01) a concentração de malondialdeído (MDA). A cafeína aumentou (P<0,05) a motilidade, a viabilidade, a PMHA e a concentração de MDA e reduziu a integridade acrossomal. Quando associados (ChA+cafeína), houve aumento (P<0,05) na motilidade, na PMHA, na viabilidade e na integridade acrossomal. Conclui-se que a adição de ChA ao meio de diluição melhora a qualidade das doses inseminantes de suínos. A adição de cafeína durante o reaquecimento só é recomendada ao sêmen adicionado de ChA quando esse for armazenado por períodos prolongados (72h), devendo a dose inseminante ser utilizada imediatamente após sua adição.(AU)

Incidence of the second parity syndrome in sows from a commercial farm / Incidência de síndrome do segundo parto em fêmeas suínas de uma granja comercial

O objetivo do presente estudo foi investigar a ocorrência da síndrome do segundo parto em uma granja comercial de suínos e apresentar alternativas para minimizar esse problema reprodutivo. Os dados foram obtidos de 363 fêmeas de genética comercial (DB-30) de primeiro e segundo partos, entre os anos de 2010 e 2011. Os animais pertenciam a uma granja comercial de ciclo completo com 1200 matrizes, cujos índices zootécnicos não permitiam detectar a presença da síndrome do segundo parto. O período de lactação foi de 24,6±3,3 dias. Foram analisados o número de nascidos totais e nascidos vivos, o peso da leitegada ao nascimento, o número de desmamados e o peso ao desmame do lote e também individualmente de cada marrã ao longo do ano. As médias e o desvio-padrão foram calculados, e os dados obtidos no primeiro e no segundo parto foram comparados pelo teste t pareado a 5%. Não houve diferença (P>0,05) no número de nascidos totais e no número de nascidos vivos entre o primeiro e o segundo parto. No entanto, constatou-se que 54% das fêmeas apresentaram igual ou menor número de nascidos no segundo parto, caracterizando a síndrome do segundo parto na maior parte dos animais. Nesse lote, o número de leitões nascidos a menos em relação ao primeiro ciclo reprodutivo foi de 3,6±2,9. Das 363 matrizes avaliadas, 153 (42%) apresentaram 16 ou mais leitões no primeiro parto. Destas, 92 (60%) tiveram menor número de leitões no segundo parto e 41 (27%) apresentaram maior número de leitões. Também se verificou maior incidência (50% ou mais) da síndrome do segundo parto nos meses de janeiro a março e de outubro a dezembro. Conclui-se que a síndrome do segundo parto é um problema que pode afetar 50% ou mais das matrizes, nem sempre detectada por meio dos índices zootécnicos da granja. Medidas como pesagem dos animais na primeira cobertura e logo após o desmame, além de programas de alimentação com dietas balanceadas, principalmente durante os meses mais quentes do ano, são ferramentas importantes para amenizar esse problema.(AU)

Técnica de levantamento de assoalho de seio maxilar associado a instalação imediata de implantes osseointegrados: relato de caso clínico / Technique of maxillary sinus floor augmentation associated with immediate installation of dental implants: a case report

A cirurgia de enxerto para levantamento de assoalho de seio maxilar é um procedimento cirúrgico previsível com importantes aplicações. Os bons resultados alcançados podem ser atribuídos a uma melhor compreensão anatômica associado a uma abordagem criteriosa na seleção de pacientes e escolha dos biomateriais seguido de um controle das possíveis complicações. O objetivo do presente estudo foi avaliar clinica e radiograficamente um caso clínico o qual foi realizado o levantamento de seio maxilar com a utilização de tecido ósseo autógeno proveniente da região de ramo mandibular e simultânea instalação de implantes osseointegrados. Pelo caso fica demonstrado que a técnica utilizada é de fácil realização e possibilita uma drástica diminuição no tempo de tratamento

The surgery of graft for maxillary sinus floor augmentation is a surgical procedure with expectation of important applications. The good results achieved can be attributed to a better understanding anatomical approach combined with a careful selection of patients and choice of biomaterials followed by a control of possible complications. The aim of this study was to evaluate clinically and radiographically a case which held the sinus lifting with the use of autogenous bone from the mandibular region and simultaneous installation of dental implants. It was demonstrated that the technique is easy and enables a drastic reduction in treatment time

Densidade de plantio e idade de colheita de quebra-pedra [Phyllanthus amarus (Schumach. & Thonning) genótipo Unicamp-CPQBA 14] na produtividade de filantina / Density of planting and age of Phillanthus amarus genotype UNICAMP/CPQBA-14 on the productivity of phyllanthin

Avaliou-se o efeito da densidade de plantio e da idade de Phyllanthus amarus CPQBA-14, sobre o teor e produtividade de filantina. O delineamento experimental foi em blocos ao acaso, em esquema fatorial 4 x 6, com cinco repetições, envolvendo 4 tratamentos de densidade de plantio (400.000 pl ha -1; 200.000 pl ha -1; 100.000 pl ha -1 e 66.667,68 pl ha -1) com 6 colheitas (30, 45, 60, 75, 90 e 105 dias após o transplante (DAT)). Os dados foram submetidos a Análise de Variâncias e Regressão (P > 0.001). Para o teor de Filantina, foram observados efeitos significativos e independentes dos tratamentos de idade da planta na colheita e densidade de plantio. O teor de filantina aumentou linearmente com a idade da planta, atingindo o máximo aos 105 DAT, com teor de 11,52 g Kg-1. O adensamento populacional estimado que proporcionou máximo teor de filantina (8,66 g kg-1) foi de 299.860 pl ha-1. A produtividade de filantina apresentou interação significativa entre densidade de plantio e idade da planta na colheita. Observou-se que ao longo do crescimento da planta, os tratamentos com 200 e 400.000 pl ha -1 apresentaram as maiores produtividades de filantina (39,3 e 37,8 kg ha-1) aos 97,12 e 95,17 DAT, respectivamente. Conclui-se que o espaçamento apropriado para o melhor arranjo espacial de cultivo é de 299.860 pl ha-1 para as produtividades de 3.974,19 Kg (massa de matéria seca foliar) x 0,00866 kg (teor de filantina) = 34,416 kg ha-1 de filantina, com colheita programada aos 97 DAT.

The effect of planting density and age of Phyllanthus amarus- genotype CPQBA-14, on the content and productivity of phyllanthin were evaluated. The experimental design was randomized blocks in factorial scheme 4 x 6, with five replicates, consisting of 4 treatments of planting density (400.000 pl ha-1; 200.000 pl ha-1; 100.000 pl ha-1 and 66667.68 pl ha-1) with six ages of plants (30, 45, 60, 75, 90 and 105 days after transplanting (DAT)). Data were subjected to analysis of variance and regression (P>0.005). For phyllanthin content independent and significant effects of harvest and planting density were observed. The phyllanthin contents increased with plant age, peaking at 105 DAT, with a content of 11.52 g kg-1. The estimated population density that provided the high phyllanthin contents (8.66 g kg-1) was at 299.860 pl ha-1. The productivity of phyllanthin showed significant interaction between planting density and plants ages. It was observed during the plant growth, the treatments with 200 and 400.000 pl ha -1 had the highest yield of phyllanthin (39.3 and 37.8 kg ha-1) with 95.17 and 97.12 DAT, respectively. We conclude that the proper spacing for best spatial arrangement of the cultivation is 299.860 pl ha-1 for the yield of 3974.19 kg (dry weight leaf) x 0.00866 kg (phyllanthin contents) = 34.416 kg ha-1 of phyllanthin with harvest scheduled at 97 DAT.

Atividade hepatoprotetora dos extratos etanólico e hexânico das folhas de Momordica charantia / Hepatoprotective activity of hexane and ethanol extracts from Momordica charantia L. leaves

Momordica charantia L. (Cucurbitaceae) é empregada na medicina popular pelas diversas propriedades biológicas e farmacológicas. Neste trabalho, objetivou-se avaliar a atividade hepatoprotetora dos extratos hexânico (EH) e etanólico (EE) das folhas de M. charantia no modelo de lesão hepática aguda induzida pelo etanol. Em ambos os extratos foram detectados esteróides; EH apresentou atividade antioxidante mais marcante que EE. O pré-tratamento de camundongos (Swiss, machos) por via oral por três dias consecutivos com 25, 50 ou 100 mg Kg-1 de EH preveniu o aumento tanto de AST quanto de ALT (p<0,05) enquanto o pré-tratamento com EE preveniu apenas o aumento de ALT (p<0,05). Ambos os extratos reduziram a atividade das enzimas hepáticas, contudo EH apresentou efeito mais relevante, caracterizando potencial hepatoprotetor, possivelmente associado ao seu potencial antioxidante.

Momordica charantia L. (Cucurbitaceae) has been used in folk medicine based on its several biological and pharmacological properties. The aim of this work was to evaluate the hepatoprotective activity of hexane (HE) and ethanol (EE) extracts from M. charantia leaves on the acute liver injury model induced by ethanol. Steroids were detected in both extracts, and EH had higher antioxidant activity than EE. Oral pretreatment of male Swiss mice for three consecutive days with 25, 50 or 100 mg kg-1 HE prevented the increase in AST and ALT (p<0.05), whereas pretreatment with EE only prevented ALT increase (p<0.05). Both extracts reduced the activity of hepatic enzymes; however, HE had a more relevant effect, characterizing hepatoprotective potential, likely associated with its antioxidant potential.

Effects of sirolimus alone or in combination with cyclosporine A on renal ischemia/reperfusion injury

Calcineurin inhibitors exacerbate ischemic injury in transplanted kidneys, but it is not known if sirolimus protects or exacerbates the transplanted kidney from ischemic injury. We determined the effects of sirolimus alone or in combination with cyclosporin A (CsA) on oxygenated and hypoxic/reoxygenated rat proximal tubules in the following in vitro groups containing 6-9 rats per group: sirolimus (10, 50, 100, 250, 500, and 1000 çg/mL); CsA (100 µg/mL); sirolimus (50 and 250 çg/mL) + CsA (100 µg/mL); control; vehicle (20 percent ethanol). For in vivo studies, 3-week-old Wistar rats (150-250 g) were submitted to left nephrectomy and 30-min renal artery clamping. Renal function and histological evaluation were performed 24 h and 7 days after ischemia (I) in five groups: sham, I, I + SRL (3 mg·kg-1·day-1, po), I + CsA (3 mg·kg-1·day-1, sc), I + SRL + CsA. Sirolimus did not injure oxygenated or hypoxic/reoxygenated proximal tubules and did not potentiate the tubular toxic effects of CsA. Neither drug affected the glomerular filtration rate (GFR) at 24 h. GFR was reduced in CsA-treated rats on day 7 (0.5 ± 0.1 mL/min) but not in rats receiving sirolimus + CsA (0.8 ± 0.1 mL/min) despite the reduction in renal blood flow (3.9 ± 0.5 mL/min). Acute tubular necrosis regeneration was similar for all groups. Sirolimus alone was not toxic and did not enhance hypoxia/reoxygenation injury or CsA toxicity to proximal tubules. Despite its hemodynamic effects, sirolimus protected post-ischemic kidneys against CsA toxicity.

Caracterización de lesiones hepáticas sólidas con técnica de difusión por resonancia magnética: reporte preliminar / Characterization of solid liver lesions with diffusion-weighted MRI technique

The characterization of liver lesions in MRI is based on morphology, behavior in the various sequences, and with paramagnetic contrast. Objective: To evaluate the diffusion behavior of various solid lesions, measuring ADC to determine its usefulness. Between 2007 and 2008 we studied 51 solid focal lesions (26 patients) with MRI, using conventional sequences and diffusion-weighted imaging obtained with EPI mode with different values of "b". Lesions corresponded to 20 hemangiomas, 12 focal nodular hyperplasia (FNH), 5 hepatocellular carcinoma (HCC), and 14 metastases. The statistical analysis allowed us to determine optimal cutoff level (ADC 1.28 x 1Cr³mm²/s) to differentiate benign from malignant lesions. ADC values of benign lesions were significantly higher than those of the malignant ones; the hemangioma was the solid lesion with higher ADC values, followed by FNH, HCC and metastases, with lower values. Registered mean values were 1.68, 1.30, 1.08 and 1.03 (x10-³mrrf/s), respectively. In our view, the diffusion-weighted technique should be used to supplement conventional MRI for proper characterization of solid liver lesions.

La caracterización de las lesiones hepáticas en resonancia magnética se basa en: morfología, comportamiento en las diversas secuencias y con el contraste paramagnético. Objetivo: evaluar el comportamiento en difusión de diversas lesiones sólidas, midiendo ADC para determinar su utilidad. En los años 2007-2008, se estudiaron 51 lesiones focales sólidas (26 pacientes) con resonancia magnética, utilizando secuencias convencionales y difusión, que fue adquirida con modalidad EPI con diferentes valores de "b". Las lesiones correspondieron a: 20 hemangiomas, 12 hiperplasias nodulares focales (HNF), 5 hepatocarcinomas (HCC) y 14 metástasis. El análisis estadístico permitió determinar nivel de corte óptimo (ADC 1.28x10'³mrrf/s) para diferenciar lesiones benignas de malignas. Los valores ADC de lesiones benignas fueron significativamente mayores que los de las malignas; el hemangioma fue la lesión sólida con mayor ADC, seguido por HNF, HCC y metástasis, con valores menores. Los valores medios encontrados fueron de 1.68, 1.30, 1.08 y 1.03 (x10³mm²/s) respectivamente. En nuestra opinión, la técnica de difusión debe utilizarse como complemento de resonancia magnética convencional, para una adecuada caracterización de lesiones sólidas hepáticas.

The association of ACE gene D/I polymorphism with cardiovascular risk factors in a population from Rio de Janeiro

Our aim was to determine the frequencies of the angiotensin-converting enzyme (ACE) gene alleles D and I and any associations to cardiovascular risk factors in a population sample from Rio de Janeiro, Brazil. Eighty-four adults were selected consecutively during a 6-month period from a cohort subgroup of a previous large cross-sectional survey in Rio de Janeiro. Anthropometric data and blood pressure measurements, echocardiogram, albuminuria, glycemia, lipid profile, and ACE genotype and serum enzyme activity were determined. The frequency of the ACE*D and I alleles in the population under study, determined by PCR, was 0.59 and 0.41, respectively, and the frequencies of the DD, DI, and II genotypes were 0.33, 0.51, and 0.16, respectively. No association between hypertension and genotype was detected using the Kruskal-Wallis method. Mean plasma ACE activity (U/mL) in the DD (N = 28), DI (N = 45) and II (N = 13) groups was 43 (in males) and 52 (in females), 37 and 39, and 22 and 27, respectively; mean microalbuminuria (mg/dL) was 1.41 and 1.6, 0.85 and 0.9, and 0.6 and 0.63, respectively; mean HDL cholesterol (mg/dL) was 40 and 43, 37 and 45, and 41 and 49, respectively, and mean glucose (mg/dL) was 93 and 108, 107 and 98, and 85 and 124, respectively. A high level of ACE activity and albuminuria, and a low level of HDL cholesterol and glucose, were found to be associated with the DD genotype. Finally, the II genotype was found to be associated with variables related to glucose intolerance.

Atividade antimicrobiana de extratos hidroalcólicos de espécies da coleção de plantas medicinais CPQBA/UNICAMP

Extratos obtidos a partir de 45 espécies da Coleção de Germoplasmas do CPQBA foram estudados quanto à atividade antimicrobiana. As espécies que apresentaram forte inibição (Concentração Mínima Inibitória até 0,5 mg/mL) para os respectivos microrganismos foram: Achillea millefolium (0,5), Mikania laevigata (0,04), Solidago chilensis (0,1), Piper marginatum (0,2) para Staphylococcus aureus; Aloysia gratissima (0,1), P. marginatum (0,2), M. laevigata (0,09) para Bacillus subtilis e Mentha pullegium (0,3), Mikania glomerata (0,1), M. laevigata (0,04), Stachytarpeta cayenensis (0,2) e Bacharis dracunculifolia (0,5) para Streptococcus faecium. De acordo com os resultados, ressaltamos a espécie M. laevigata por apresentar inibição contra três das bactérias estudadas, em concentrações similares a do cloranfenicol, padrão de referência utilizado.

Crude extracts of 45 medicinal plants from CPQBA Germoplasm Collection were tested for their antimicrobial activity. The species that presented strong activity (Minimal Inhibitory Concentration until 0.5 mg.mL-1) against the respective microorganisms were: Achillea millefolium (0.5), Mikania laevigata (0.04), Solidago chilensis (0.1), Piper marginatum (0.2) against Staphylococcus aureus; Aloysia gratissima (0.1), P. marginatum (0.2), M. laevigata (0.09) against Bacillus subtilis and Mentha pullegium (0.3), Mikania glomerata (0.1), M. laevigata (0.04), Stachytarpeta cayenensis (0.2) and Bacharis dracunculifolia (0.5) against Streptococcus faecium. According to results, M. laevigata was able to inhibit three bacteria species, at similar concentrations as observed for the standard chloranphenicol.

Adhesion of the human pathogen Sporothrix schenckii to several extracellular matrix proteins

The pathogenic fungus Sporothrix schenckii is the causative agent of sporotrichosis. This subcutaneous mycosis may disseminate in immunocompromised individuals and also affect several internal organs and tissues, most commonly the bone, joints and lung. Since adhesion is the first step involved with the dissemination of pathogens in the host, we have studied the interaction between S. schenckii and several extracellular matrix (ECM) proteins. The binding of two morphological phases of S. schenckii, yeast cells and conidia, to immobilized type II collagen, laminin, fibronectin, fibrinogen and thrombospondin was investigated. Poly (2-hydroxyethyl methacrylate) (poly-HEMA) was used as the negative control. Cell adhesion was assessed by ELISA with a rabbit anti-S. schenckii antiserum. The results indicate that both morphological phases of this fungus can bind significantly to type II collagen, fibronectin and laminin in comparison to the binding observed with BSA (used as blocking agent). The adhesion rate observed with the ECM proteins (type II collagen, fibronectin and laminin) was statistically significant (P<0.05) when compared to the adhesion obtained with BSA. No significant binding of conidia was observed to either fibrinogen or thrombospondin, but yeast cells did bind to the fibrinogen. Our results indicate that S. schenckii can bind to fibronectin, laminin and type II collagen and also show differences in binding capacity according to the morphological form of the fungus

Fomes pectinatis: an aeroallergen in India.

This work is focused on the aerobiology and allergenicity of Fomes pectinatis in India. The atmospheric concentration of Fomes basidiospores was recorded and the antigens were prepared from spore (FSE) and whole body (FWBE) materials. The intradermal (ID) and prick (PT) skin tests were conducted on 172 patients having respiratory allergy. The period from July to October has been recorded as having a higher concentration of Fomes spores. The maximum counts (67 spores/m3) were observed from the North Delhi site in the month of July, 1989, compared with 550 spores/m3 in the South Delhi site. Marked skin positivity (2+ and above) varied from 9.8% to FSE to 22% to FWBE. Nine out of twelve ID positive patients (2+ to 3+) to FSE also gave PT positive response. For FWBE, similar ID and PT response was obtained in 80% of cases. The soluble protein content of FSE was 0.37 mg/ml, whereas, for FWBE it was 0.70 mg/ml. It was observed through ELISA that almost all patients had significantly raised FP specific IgE levels in their sera. The current study, therefore, indicates that Fomes pectinatis may be a prevalent aeroallergen in India.

Changes in the TBARs content and superoxide dismutase, catalase and glutathione peroxidase activities in the lymphoid organs and skeletal muscles of adrenodemedullated rats

Thiobarbituric acid reactant substances (TBARs) content, and the activities of glucose-6-phosphate dehydrogenase (G6PDh), citrate synthase (CS), Cu/Zn- and Mn-superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPX) were measured in the lymphoid organs (thymus, spleen, and mesenteric lymph nodes (MLN)) and skeletal muscles (gastrocnemius and soleus) of adrenodemedullated (ADM) rats. The results were compared with those obtained for sham-operated rats. TBARs content was reduced by adrenodemedullation in the lymphoid organs (MLN (28 percent), thymus (40 percent) and spleen (42 percent)) and gastrocnemius muscle (67 percent). G6PDh activity was enhanced in the MLN (69 percent) and reduced in the spleen (28 percent) and soleus muscle (75 percent). CS activity was reduced in all tissues (MLN (75 percent), spleen (71 percent), gastrocnemius (61 percent) and soleus (43 percent)), except in the thymus which displayed an increment of 56 percent. Cu/Zn-SOD activity was increased in the MLN (126 percent), thymus (223 percent), spleen (80 percent) and gastrocnemius muscle (360 percent) and was reduced in the soleus muscle (31 percent). Mn-SOD activity was decreased in the MLN (67 percent) and spleen (26 percent) and increased in the thymus (142 percent), whereas catalase activity was reduced in the MLN (76 percent), thymus (54 percent) and soleus muscle (47 percent). It is particularly noteworthy that in ADM rats the activity of glutathione peroxidase was not detectable by the method used. These data are consistent with the possibility that epinephrine might play a role in the oxidative stress of the lymphoid organs. Whether this fact represents an important mechanism for the establishment of impaired immune function during stress remains to be elucidated

Rejuvenation of epididymal principal cells by prolactin under androgen deficiency.

Influence of prolactin on the ultrastructure of principal cells lining the epididymal epithelium was investigated in Wistar rats. Orchidectomy produced degenerative changes suggesting that structural integrity of principal cell is maintained by factors originating in the testis. The atrophic changes in the principal cell of ordhidectomised rats were significantly reversed when prolactin was administered to these animals. The number of cells that responded were found to increase with the dose of prolactin injected. On the otherhand, bromocryptine treatment did not appreciably change the ultrastructure of principal cells in orchidectomised rats. Results suggest that prolactin may have a rejuvenating epididymal principal cells in androgen deficient states.

Impact of prolactin on epididymal lipid profile in castrated rats.

Prolactin treatment to castrated rats led to accumulation of triacylglycerol and esterified cholesterol. There was no appreciable drift in epididymal cholesterol: phospholipid ratio between the prolactin treated and control animals. However, further analysis of phospholipids showed a build up of phosphatidyl inositol, phosphatidyl choline and phosphatidyl ethanolamine but a drop in the levels of phosphatidyl serine and sphingomyelin in prolactin treated castrated rats as compared to those castrated animals injected with vehicle alone. Changes in phospholipids reported above were prominently seen in the group of castrated rats that received 100 micrograms oPRL/100 g body weight but not in those animals which received either lower or higher doses of the hormone. Interestingly, bromocryptine treatment in castrated rats produced a general depletion in the levels of all lipid classes studied in the epididymis. It is suggested that this may be due to impaired synthesis and/or increased breakdown of lipids in this organ.

Survey of airborne culturable and non-culturable fungi at different sites in Delhi metropolis.

A two year aerobiological survey for culturable and non-culturable fungi was conducted at human height at five different sites in Delhi metropolis. Burkard Personal Volumetric Sampler for petriplates and slide exposures were used for sampling the air. With simultaneous petriplate and slide exposure a total of 98 fungal forms were recorded. Cladosporium contributed for 25-40% of total airborne fungi followed by Ustilago (smuts) (24%) Aspergillus flavus (10-13%), Alternaria (11%) and A. niger (8%). Basidiomycetes contributed 7-13% at different sites. The frequency of occurrence of these types varied from 50-98%. In general fungal concentration was high from July to April with low counts in winter (January) and dry and hot summer (May-June). Quantitative variations in the spore counts were found to be statistically significant within the same urban locality.

Modulation in activity of some epididymal glycosidases by prolactin.

Activity of glycosidases in the epididymis was influenced by several factors originating in the testis. Activities of all the three glycosidases studied viz., beta-glucosidase, beta-galactosidase and alpha-mannosidase were found to be significantly lower in the epididymis of orchidectomized animals than in sham operated rats. However, an enhanced activity of epididymal beta-galactosidase and alpha-mannosidase was noticed in prolactin treated orchidectomized rats compared to orchidectomized rats given vehicle alone. On the other hand, activity of these two enzymes in bromocriptine treated orchidectomized rats was even lower than that found in orchidectomized rats given vehicle. Neither prolactin nor bromocriptine treatment had any significant influence on the epididymal beta-glucosidase. The results suggest a selective but definite action of prolactin on epididymal glycosidases.

Ultrasonographic localization for renal biopsy.

One hundred and fifty consecutive percutaneous renal biopsies were performed using ultrasonography to localise the site and depth of the lower pole of the left kidney. Renal tissue was obtained in 95% of cases and an accurate histopathological diagnosis was reached in 89% of patients. Gross haematuria following the procedure occurred in 6%, but was usually transient. No other complications were encountered. Ultrasonographic marking of the biopsy site and depth is a quick, simple, accurate and safe method of localizing the kidney for the purpose of a biopsy.